Compounds useful for the treatment of diseases responsive to antiangiogenetic therapy

ABSTRACT

This invention relates to the use of certain compounds for the treatment of diseases that are responsive to antiangiogenetic therapy. Excessive angiogenesis occurs in diseases such as cancer, diabetic blindness, age-related macular degeneration, rheumatoid arthritis and psoriasis. The compounds are particularly useful for anti-metastatic treatment or for the treatment of age-related macular degeneration.

TECHNICAL FIELD

This invention relates to the use of certain compounds for the treatmentof diseases that are responsive to antiangiogenetic therapy, inparticular for anti-metastatic treatment or for the treatment ofage-related macular degeneration.

BACKGROUND ART

Angiogenesis (or neovascularisation) is the formation of new bloodvessels by sprouting from preexisting vessels. Angiogenesis is generallyabsent in healthy adult or mature tissue. However, it occurs in thehealthy body for healing wounds and for restoring blood flow to tissuesafter injury or insult. In females, angiogenesis also occurs during themonthly reproductive cycle and during pregnancy. Under these processes,the formation of new blood vessels is strictly regulated.

In many serious disease states, the body loses control overangiogenesis. Excessive angiogenesis occurs in diseases such as cancer,diabetic blindness, age-related macular degeneration, rheumatoicarthritis, and psoriasis. In these conditions, new blood vessels feeddiseased tissues, destroy normal tissues, and in the case of cancer, thenew vessels allow tumor cells to escape into the circulation and lodgein other organs (tumor metastasis).

Experimental evidence has accumulated over the years to show that avariety of strategies that limit angiogenesis also slow or inhibittumour growth, suggesting that blocking tumour-induced angiogenesis is avalid, novel approach to tumour therapy.

Age-related macular degeneration (AMD) is a common eye disease thatgradually destroys central visual function; AMD occurs in stages fromthe early stage dry form to the later-stage and more severe wet formassociated with the formation of new abnormal blood vessels in the backof the eye.

Thus there is a continued need for new antiangiogenetic therapies aimingat halting new blood vessel growth.

WO 98/47879 and WO 00/24707 (NeuroSearch A/S) describe a number ofsubstituted phenyl derivatives active as chloride channel blockers.

WO 00/76495 (Smithkline Beecham Corp.) describes a number of substitutedphenyl derivatives active as IL-8 receptor antagonists.

SUMMARY OF THE INVENTION

It is an object of the invention to provide new therapies for treatingdiseases that are responsive to antiangiogenetic therapy. In particular,it is an object to provide therapies for arresting tumor growth andpreventing the formation of metastases. A further object of theinvention is the provision of therapies for treating age-related maculardegeneration.

In its first aspect, the invention provides the use of a compound ofgeneral formula I

or a pharmaceutically acceptable salt thereof for the manufacture of apharmaceutical composition for the treatment, prevention or alleviationof a disease or a disorder or a condition of a mammal, including ahuman, which disease, disorder or condition is responsive to inhibitionof angiogenesis.

In a second aspect, the invention provides the use of a VRAC blocker ora pharmaceutically acceptable salt thereof for the manufacture of apharmaceutical composition for the treatment, prevention or alleviationof age-related macular degeneration.

Other objects of the invention will be apparent to the person skilled inthe art from the following detailed description and examples.

DETAILED DISCLOSURE OF THE INVENTION

According to the invention it has now been found that certain compoundscan be used for the treatment of diseases that are responsive toantiangiogenetic therapy, in particular for anti-metastatic treatment.

Thus, in its first aspect, the invention relates to the use of acompound of general formula I

or a pharmaceutically acceptable salt thereof

-   wherein R² represents tetrazolyl; and    -   R³, R⁴, R⁵, R⁶, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ independently of        each other represent hydrogen, halo, trifluoromethyl, nitro,        alkyl, alkylcarbonyl, —NR^(a)R^(b), —NR^(a)—CO—R^(b), phenyl or        heteroaryl;    -   which phenyl is optionally substituted with halo,        trifluoromethyl, nitro, —CO—NHR^(c), —CO—O—R^(c) or —CO—NR′R″;    -   wherein R^(c) is hydrogen, alkyl, or phenyl;    -   R′ and ″ independently of each other are hydrogen or alkyl; or    -   R′ and R″ together with the nitrogen to which they are attached        form a 5- to 7-membered heterocyclic ring, which ring may        optionally comprise as a ring member, one oxygen atom, and/or        one additional nitrogen atom, and/or one carbon-carbon double        bond, and/or one carbon-nitrogen double bond;    -   and which heterocyclic ring may optionally be substituted with        alkyl;    -   R^(a) and R^(b) independently of each other are hydrogen or        alkyl; or    -   R¹⁵ and R¹⁶, or R¹⁴ and R¹⁵ together with the phenyl ring to        which they are attached form a naphthyl ring or an indanyl ring;        and R³, R⁴, R⁵, R⁶, R¹² and R¹³ and the remaining one of R¹⁴,        R¹⁵ and R¹⁶ are as defined above;        for the manufacture of a pharmaceutical composition for the        treatment, prevention or alleviation of a disease or a disorder        or a condition of a mammal, including a human, which disease,        disorder or condition is responsive to inhibition of        angiogenesis.

In a special embodiment, the invention relates to the use of a compoundof general formula I

or a pharmaceutically acceptable salt thereof

-   wherein R² represents tetrazolyl;-   R³, R⁴, R⁵, R⁶, R¹², R¹³, R⁴, R¹⁵, and R¹⁶ independently of each    other represent hydrogen halo, trifluoromethyl, nitro, or phenyl;-   which phenyl is optionally substituted with halo, trifluoromethyl,    nitro, or —CO—NHR^(c);-   wherein R^(c) is hydrogen, alkyl, or phenyl;-   for the manufacture of a pharmaceutical composition for the    treatment, prevention or alleviation of a disease or a disorder or a    condition of a mammal, including a human, which disease, disorder or    condition is responsive to inhibition of angiogenesis.

In another aspect, the invention relates to a method of treatment,prevention or alleviation of a disease or a disorder or a condition of aliving animal body, including a human, which disorder, disease orcondition is responsive to inhibition of angiogenesis, comprising thestep of administering to such a living animal body, including a human,in need thereof a therapeutically effective amount of a compound ofgeneral formula I

or a pharmaceutically acceptable salt thereof

-   wherein R² represents tetrazolyl; and    -   R³, R⁴, R⁵, R⁶, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ independently of        each other represent hydrogen, halo, trifluoromethyl, nitro,        alkyl, alkylcarbonyl, —NR^(a)R^(b), —NR^(a)—CO—R^(b), phenyl or        heteroaryl;    -   which phenyl is optionally substituted with halo,        trifluoromethyl, nitro, —CO—NHR^(c), —CO—O—R^(c) or —CO—NR′R″;    -   wherein R^(c) is hydrogen, alkyl, or phenyl;    -   R′ and R″ independently of each other are hydrogen or alkyl; or    -   R′ and R″ together with the nitrogen to which they are attached        form a 5 to 7-membered heterocyclic ring, which ring may        optionally comprise as a ring member, one oxygen atom, and/or        one additional nitrogen atom, and/or one carbon-carbon double        bond, and/or one carbon-nitrogen double bond;    -   and which heterocyclic ring may optionally be substituted with        alkyl;    -   R^(a) and R^(b) independently of each other are hydrogen or        alkyl or    -   R¹⁵ and R¹⁶, or R¹⁴ and R¹⁵ together with the phenyl ring to        which they are attached form a naphthyl ring or an indanyl ring;        and R³, R⁴, R⁵, R⁶, R¹² and R¹³ and the remaining one of R¹⁴,        R¹⁵ and R¹⁶ are as defined above.

In a special embodiment, the invention relates to a method of treatment,prevention or alleviation of a disease or a disorder or a condition of aliving animal body, including a human, which disorder, disease orcondition is responsive to inhibition of angiogenesis, comprising thestep of administering to such a living animal body, including a human,in need thereof a therapeutically effective amount of a compound ofgeneral formula I

or a pharmaceutically acceptable salt thereof

-   wherein R² represents tetrazolyl;-   R³, R⁴, R⁵, R⁶, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ independently of each    other represent hydrogen, halo, trifluoromethyl, nitro, or phenyl;-   which phenyl is optionally substituted with halo, trifluoromethyl,    nitro, or —CO—NHR^(c);-   wherein R^(c) is hydrogen, alkyl, or phenyl.

The living animal body to be treated according to this invention ispreferably a mammal, most preferably a human, in need for suchtreatment.

In one embodiment of the compound of general formula I, R² representstetrazolyl; R³, R⁴, R⁵, R⁶, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ independently ofeach other represent hydrogen, halo, trifluoromethyl, or nitro.

In a second embodiment of the compound of general formula I, R³, R⁵, andR⁶ represent hydrogen; and R⁴ represents halo, such as bromine.

In a third embodiment of the compound of general formula I, R³, R4, R⁵and R⁶ represent hydrogen.

In a further embodiment of the compound of general formula I, R³, R⁵,and R⁶ represent hydrogen; and R⁴ represents —NR^(a)R^(b), such asamino.

In a still further embodiment of the compound of general formula I, R³,R⁵, and R⁶ represent hydrogen; and R⁴ represents —NR^(a)—CO—R^(b), suchas acetylamino.

In a further embodiment of the compound of general formula I, R³, R⁵,and R⁶ represent hydrogen; and R⁴ represents phenyl substituted withtrifluoromethyl, nitro or —CO—NHR^(c); wherein R^(c) is phenyl. In aspecial embodiment, R⁴ represents phenyl substituted withtrifluoromethyl, such as 4-trifluoromethylphenyl. In a furtherembodiment, R⁴ represents phenyl substituted with nitro, such as3-nitrophenyl. In a further embodiment, R⁴ represents phenyl substitutedwith —CO—NHR^(c), such as anilinocarbonylphenyl, in particular4-anilinocarbonylphenyl.

In a further embodiment of the compound of general formula I, R³, R⁵,and R⁶ represent hydrogen; and R⁴ represents phenyl substituted with—CO—O—R^(c) or —CO—NR′R″. In a special embodiment, R⁴ represents phenylsubstituted with —CO—O—R^(c), wherein R^(c) is hydrogen. In a specialembodiment, R⁴ represents phenyl substituted with —CO—NR′R″, such as4-dimethylcarbamoylphenyl or 4-(4-methyl-1-piperazine-carbonyl)-phenyl.

In a still further embodiment, R¹⁵ represents trifluoromethyl. In aspecial embodiment, R¹⁵ represents trifluoromethyl and R¹², R¹³, R¹⁴ andR¹⁶ represent hydrogen. In a further embodiment R¹³ representstrifluoromethyl. In a special embodiment, R¹³ and R¹⁵ representtrifluoromethyl and R¹², R¹⁴ and R¹⁶ represent hydrogen.

In a further embodiment, R¹⁶ represents trifluoromethyl. In a specialembodiment, R¹⁶ represents trifluoromethyl and R¹², R¹³, R¹⁴ and R¹⁵represent hydrogen.

In a still further embodiment, R¹⁵ represents halo, such as chloro orbromo. In a special embodiment, R¹⁵ represents halo, such as chloro orbromo, and R¹², R¹³, R¹⁴ and R¹⁶ represent hydrogen. In a furtherembodiment, R¹³ and R¹⁵ represent halo, such as chloro, and R¹², R¹⁴ andR¹⁶ represent hydrogen. In a still further embodiment, R¹⁴ and R¹⁵represent halo, such as chloro, and R¹², R¹³ and R¹⁶ represent hydrogen.

In a further embodiment, R¹⁶ represents halo, such as fluoro. In aspecial embodiment, R¹⁶ represents halo, such as fluoro, and R¹², R¹³,R¹⁴ and R¹⁵ represent hydrogen.

In a still further embodiment, R¹⁶ represents alkyl, such as methyl orethyl. In a special embodiment, R¹⁶ represents alkyl, such as methyl orethyl, and R¹², R¹³, R¹⁴ and R¹⁵ represent hydrogen.

In a further embodiment, R¹⁴ represents nitro. In a special embodiment,R¹⁴ represents nitro, and R¹², R¹³, R¹⁵ and R¹⁶ represent hydrogen.

In a still further embodiment, R¹⁴ represents alkylcarbonyl, such asacetyl. In a special embodiment, R¹⁴ represents alkylcarbonyl, such asacetyl, and R¹², R¹³ , R¹⁵ and R¹⁶ represent hydrogen.

In a further embodiment, R¹⁵ represents phenyl. In a further embodiment,R¹⁴ represents phenyl. In a special embodiment, one of R¹⁴ or R¹⁵represents phenyl, and the remaining of R¹², R¹³, R¹⁴, R¹⁵ and R¹⁶represent hydrogen.

In a still further embodiment, R¹⁵ represents pyridyl, such aspyridin-3-yl. In a special embodiment, R¹⁵ represents pyridyl, such aspyridin-3-yl, and R¹², R¹³, R¹⁴ and R¹⁶ represent hydrogen.

In a further embodiment, R¹⁵ and R¹⁶ together with the phenyl ring towhich they are attached form a naphthyl ring.

In a still further embodiment, R¹⁴ and R¹⁵ together with the phenyl ringto which they are attached form an indanyl ring.

In a further embodiment, the compound of general formula I is

-   N-4-Nitrophenyl-N′-[4-bromo-2-(1-H-tetrazol-5-yl)phenyl]urea;-   N-3,5-Di(trifluoromethyl)phenyl-N′-[4-bromo-2-(1-H-tetrazol-5-yl)phenyl]urea;-   N-3-Trifluoromethylphenyl-N′-[4-(3-nitrophenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;-   N-3-Trifluoromethylphenyl-N′-[4-(4-anilinocarbonylphenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;-   N-3-Trifluoromethylphenyl-N′-[4-(4-trifluoromethylphenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;-   N-(3-Trifluoromethyl-phenyl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Trifluoromethyl-phenyl)-N′-[4-bromo-2-(1-H-terazol-5-yl)-phenyl]urea;-   N-(3-Trilfuoromethyl-phenyl)-N′-[4-phenyl-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Chloro-phenyl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Trifluoromethyl-phenyl)-N′-[4-amino-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Trifluoromethyl-phenyl)-N′-[4-acetylamino-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Trilfuoromethyl-phenyl)-N′-[4-carbamoyl-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Trifluoromethyl-phenyl)-N′-(4-(N″,N″-dimethylcarbamoyl)-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   3′-(1-H-tetrazol-5-yl)-4′-[3-(3-trifluoromethyl-phenyl)-ureido]-biphenyl-4-carboxylic    acid;-   N-(Indan-5-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(Biphenyl-4-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(Biphenyl-3-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Acetyl-phenyl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(Biphenyl-3-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-[3-(Pyridin-3-yl)-phenyl]-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Bromo-phenyl)-N′-[4′-(4-methyl-piperazine-1-carbonyl)-3-(1-H-tetrazol-5-yl)-biphenyl-4-yl]urea;-   N-(3,5-Dichloro-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3,4-Dichloro-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(Naphthalen-1-yl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(2-Trifluoromethyl-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(2-Fluoro-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(2-Ethyl-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   or a pharmaceutically acceptable salt thereof.

In a still further embodiment, the disease, disorder or condition thatis responsive to inhibition of angiogenesis is selected from the groupconsisting of cancer, prostate cancer, lung cancer, breast cancer,bladder cancer, renal cancer, colon cancer, gastric cancer, pancreaticcancer, ovarian cancer, melanoma, hepatoma, sarcoma, lymphoma, exudativemacular degeneration, age-related macular degeneration, retinopathy,diabetic, proliferative diabetic retinopathy, diabetic macular edema(DME), ischemic retinopathy, retinopathy of prematurity, neovascularglaucoma, corneal neovascularization, rheumatoid arthritis, andpsoriasis.

In a special embodiment of the invention, the compound is

-   N-4-Nitrophenyl-N′-[4-bromo-2-(1-H-tetrazol-5-yl)phenyl]urea;-   N-3,5-Di(trifluoromethyl)phenyl-N-[4-bromo-2-(1-H-tetrazol-5-yl)phenyl]urea;-   N-3-Trifluoromethylphenyl-N′-[4-(3-nitrophenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;-   N-3-Trifluoromethylphenyl-N′-[4-(4-anilinocarbonylphenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;-   N-3-Trifluoromethylphenyl-N′-[4-(4-trifluoromethylphenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;-   or a pharmaceutically acceptable salt thereof, and-   the treatment is an anti-metastatic treatment.

In a further aspect, the invention relates to the use of a VRAC blockeror a pharmaceutically acceptable salt thereof for the manufacture of apharmaceutical composition for the treatment, prevention or alleviationof age-related macular degeneration of a mammal, including a human.

In a still further aspect the invention relates to a method oftreatment, prevention or alleviation of age-related macular degenerationof a living animal body, including a human comprising the step ofadministering to such a living animal body, including a human, in needthereof a therapeutically effective amount of a VRAC blocker or apharmaceutically acceptable salt thereof.

In one embodiment, the VRAC blocker is a compound of general formula I

or a pharmaceutically acceptable salt thereof

-   wherein R² represents tetrazolyl; and    -   R³, R⁴, R⁵, R⁶, R¹² , R¹³, R¹⁴, R¹⁵, and R¹⁶ independently of        each other represent hydrogen, halo, trifluoromethyl, nitro,        alkyl, alkylcarbonyl, —NR^(a)R^(b), —NR^(a)—CO—R^(b), phenyl or        heteroaryl;    -   which phenyl is optionally substituted with halo,        trifluoromethyl, nitro, —CO—NHR^(c), —CO—O—R^(c) or —CO—NR′R″;    -   wherein R^(c) is hydrogen, alkyl, or phenyl;    -   R′ and R″ independently of each other are hydrogen or alkyl; or    -   R′ and R″ together with the nitrogen to which they are attached        form a 5- to 7-membered heterocyclic ring, which ring may        optionally comprise as a ring member, one oxygen atom, and/or        one additional nitrogen atom, and/or one carbon-carbon double        bond, and/or one carbon-nitrogen double bond;    -   and which heterocyclic ring may optionally be substituted with        alkyl;    -   R^(a) and R^(b) independently of each other are hydrogen or        alkyl; or    -   R¹⁵ and R¹⁶, or R¹⁴ and R¹⁵ together with the phenyl ring to        which they are attached form a naphthyl ring or an indanyl ring;        and R³, R⁴, R⁵, R⁶, R¹² and R¹³ and the remaining one of R¹⁴,        R¹⁵ and R¹⁶ are as defined above;        for the manufacture of a pharmaceutical composition for the        treatment, prevention or alleviation of a disease or a disorder        or a condition of a mammal, including a human, which disease,        disorder or condition is responsive to inhibition of        angiogenesis.

In a further embodiment, the VRAC blocker is a compound of generalformula I

or a pharmaceutically acceptable salt thereof

-   wherein R² represents tetrazolyl;-   R³, R⁴, R⁵, R⁶, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ independently of each    other represent hydrogen, halo, trifluoromethyl, nitro, or phenyl;-   which phenyl is optionally substituted with halo, trifluoromethyl,    nitro, or —CO—NHR^(a);-   wherein R^(a) is hydrogen, alkyl, or phenyl.

In a special embodiment, the VRAC blocker is

-   N-4-Nitrophenyl-N′-[4-bromo-2-(1-H-tetrazol-5-yl)phenyl]urea;-   N-3,5-Di(trifluoromethyl)phenyl-N′-(4-bromo-2-(1-H-tetrazol-5yl)phenyl]urea;-   N-3-Trifluoromethylphenyl-N′-[4-(3-nitrophenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;-   N-3-Trifluoromethylphenyl-N′-[4-(4-anilinocarbonylphenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;-   N-3-Trifluoromethylphenyl-N′-[4-(4-trifluoromethylphenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;-   N-(3-Trifluoromethyl-phenyl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Trifluoromethyl-phenyl)-N′-[4-bromo-2-(4-H-terazol-5-yl)-phenyl]urea;-   N-(3-Trifluoromethyl-phenyl)-N′-[4-phenyl-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Chloro-phenyl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Trifluoromethyl-phenyl)-N′-[4-amino-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Trifluoromethyl-phenyl)-N′-[4-acetylamino-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Trifluoromethyl-phenyl)-N′-[4-carbamoyl-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Trifluoromethyl-phenyl)-N′-[4-(N″,N″-dimethylcarbamoyl)-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   3′-(1-H-tetrazol-5-yl)-4′-[3-(3-trifluoromethyl-phenyl)-ureido]-biphenyl-4-carboxylic    acid;-   N-(Indan-5-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(Biphenyl-4-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(Biphenyl-3-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Acetyl-phenyl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(Biphenyl-3-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-[3-(Pyridin-3-yl)-phenyl]-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3-Bromo-phenyl)-N′-[4′-(4-methyl-piperazine-1-carbonyl)-3-(1-H-tetrazol-5-yl)-biphenyl-4-yl]urea;-   N-(3,5-Dichloro-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(3,4-Dichloro-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(Naphthalen-1-yl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(2-Trifluoromethyl-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(2-Fluoro-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;-   N-(2-Ethyl-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5yl)-phenyl]urea;-   or a pharmaceutically acceptable salt thereof.

DEFINITION OF SUBSTITUENTS

In the context of this invention halo represents fluoro, chloro, bromoor iodo.

In the context of this invention an alkyl group designates a univalentsaturated, straight or branched hydrocarbon chain. The hydrocarbon chainpreferably contain of from one to six carbon atoms (C₁₋₆-alkyl),including pentyl, isopentyl, neopentyl, tertiary pentyl, hexyl andisohexyl. In one embodiment alkyl represents a C₁₋₄alkyl group,including butyl, isobutyl, secondary butyl, and tertiary butyl. Inanother embodiment of this invention alkyl represents a C₁₋₃-alkylgroup, which may in particular be methyl, ethyl, propyl or isopropyl.

In the context of this invention a heteroaryl group designates anaromatic mono-, bi- or poly-heterocyclic group, which holds one or moreheteroatoms in its ring structure. Preferred heteroatoms includenitrogen (N), oxygen (O), and sulphur (S). Preferred monocyclicheteroaryl groups of the invention include aromatic 5- and 6 memberedheterocyclic monocyclic groups, including furanyl, in particular 2- or3-furanyl; thienyl, in particular 2 or 3-thienyl; pyrrolyl (azolyl), inparticular 1,2 or 3-pyrrolyl; oxazolyl, in particular oxazol-2,4 or5-yl; thiazolyl, in particular thiazol-2,4 or 5-yl; imidazolyl, inparticular 1,2 or 4-imidazolyl; pyrazolyl, in particular 1,3 or4-pyrazolyl; isoxazolyl, in particular isoxazol-3,4 or 5-yl;isothiazolyl, in particular isothiazol-3,4 or 5-yl; oxadiazolyl, inparticular 1,2,3-, 1,2,4-, 1,2,5- or 1,3,4-oxadiazol-3,4 or 5-yl;triazolyl, in particular 1,2,3-, 1,2,4-, 2,1,3- or 4,1,2-triazolyl;thiadiazolyl, in particular thiadiazol-3,4 or 5-yl; pyridinyl, inparticular 2,3 or 4-pyridinyl; pyridazinyl, in particular 3 or4-pyridazinyl; pyrimidinyl, in particular 2,4 or 5-pyrimidinyl;pyrazinyl, in particular 2 or 3-pyrazinyl; and triazinyl, in particular1,2, 3-, 1,2,4- or 1,3,5-triazinyl.

5- to 7-membered heterocyclic rings comprising one nitrogen atom includefor example, but not limited to, pyrolidine, piperidine, homopiperidine,pyrroline, tetrahydro-pyridine, pyrazolidine, imidazolidine, piperazine,homopiperazine, and morpholine.

VRAC

Volume regulated anion channels (VRAC) are present in most mammaliancells. An important function of VRAC is cell volume regulation: Uponswelling of the cell in hypotonic solution these channels are activatedand chloride ions flow out of the cell in parallel with potassium ions(via potassium channels) and water, thereby restoring the original cellvolume.

Although VRAC has not been cloned and therefore cannot yet be defined bytheir gene sequences, there is a number of defining characteristics forthese channels as observed with the whole cell patch clamp technique:VRAC is activated by cell swelling (hypotonic extracellular—orhypertonic intracellular media) and an important trigger for activationis lowered intracellular ionic strength rather than volume per se.Activation of VRAC is obligatory dependent on the presence ofintracellular ATP and complex intra-cellular signalling cascadesinvolving protein kinases support the activation process. VRAC. is anvoltage independent, outward rectifying anion selective channelexhibiting type I Eisenman halide selectivity sequence (implying forexample that iodide is more permeable than chloride) and having a widepore that also allows the permeation of a large number of negativelycharged or neutral organic molecules. VRAC is always blockedvoltage-dependently by DIDS, and NPPB and tamoxifen are voltageindependent blockers of this channel.

VRAC Blockers

A VRAC blocker is a compound that inhibits the transmembrane transportof chloride or any other anion or neutral molerule in response to cellswelling or decrease in intracellular ionic strength.

The potential of a given substance to act as a VRAC blocker may bedetermined using standard laboratory test methods, such as

-   a) whole cell or single channel patch clamp technology,-   b) microelectrode electrophysiology (penetrating, sharp electrodes),-   c) flux assays (e.g. radioactive as well as non-radioactive    isotopes),-   d) fluorescent dyes (e.g. membrane potential—or chloride    concentration indicators),-   e) cell volume measurements (e.g. light scattering, coulter counter    measurements, or optical methods such as image analysis)

The VRAC blocker may in particular be a diphenyl urea derivative such asthose disclosed in described in WO 98/47879 or WO 00/24707 (NeuroSearchA/S).

In one embodiment, the VRAC blockers show IC₅₀-values of less than 10μM, preferably less than 1000 nM, more preferably less than 100 nM, evenmore preferably less than 50 nM, and most preferred less than 10 nM forin vitro inhibition according to the standard test methods.

In a second embodiment the VRAC blockers show ED₅₀ values of less than50 mg/kg, preferably less than 10 mg/kg, more preferably less than 5mg/kg in the standard in vivo angiogenesis models.

Methods of Preparation

The compounds for use according to the invention may be prepared byconventional methods for chemical synthesis, e.g. those described in WO98/47879 or WO 00/24707 (NeuroSearch A/S).

Pharmaceutically Acceptable Salts

The active compound for use according to the invention may be providedin any form suitable for the intended administration. Suitable formsinclude pharmaceutically (i.e. physiologically) acceptable salts, andpre- or prodrug forms of the chemical compound of the invention.

Examples of pharmaceutically acceptable addition salts include, withoutlimitation, the non-toxic inorganic and organic acid addition salts suchas the hydrochloride derived from hydrochloric acid, the hydrobromidederived from hydrobromic acid, the nitrate derived from nitric acid, theperchlorate derived from perchloric acid, the phosphate derived fromphosphoric acid, the sulphate derived from sulphuric acid, the formatederived from formic acid, the acetate derived from acetic acid, theaconate derived from aconitic acid, the ascorbate derived from ascorbicacid, the benzenesulphonate derived from benzensulphonic acid, thebenzoate derived from benzoic acid, the cinnamate derived from cinnamicacid, the citrate derived from citric acid, the embonate derived fromembonic acid, the enantate derived from enanthic acid, the fumaratederived from fumaric acid, the glutamate derived from glutamic acid, theglycolate derived from glycolic acid, the lactate derived from lacticacid, the maleate derived from maleic acid, the malonate derived frommalonic acid, the mandelate derived from mandelic acid, themethanesulphonate derived from methane sulphonic acid, thenaphthalene-2-sulphonate derived from naphtalene-2-sulphonic acid, thephthalate derived from phthalic acid, the salicylate derived fromsalicylic acid, the sorbate derived from sorbic acid, the stearatederived from stearic acid, the succinate derived from succinic acid, thetartrate derived from tartaric acid, the toluene-p-sulphonate derivedfrom p-toluene sulphonic acid, and the like. Such salts may be formed byprocedures well known and described in the art.

Other acids such as oxalic acid, which may not be consideredpharmaceutically acceptable, may be useful in the preparation of saltsuseful as intermediates in obtaining a chemical compound for useaccording to the invention and its pharmaceutically acceptable acidaddition salt.

Examples of pharmaceutically acceptable cationic salts of a chemicalcompound of the invention include, without limitation, the sodium, thepotassium, the calcium, the magnesium, the zinc, the aluminium, thelithium, the choline, the lysine, and the ammonium salt, and the like,of a chemical compound of the invention containing an anionic group.Such cationic salts may be formed by procedures well known and describedin the art.

In the context of this invention the “onium salts” of N-containingcompounds are also contemplated as pharmaceutically acceptable salts(aza-onium salts). Preferred azaonium salts include the alkyl-oniumsalts, in particular the methyl- and the ethyl-onium salts; thecycloalkyl-onium salts, in particular the cyclopropyl-onium salts; andthe cycloalkylalkyl-onium salts, in particular thecyclopropyl-methyl-onium salts.

Pharmaceutical Compositions

While the active compound for use in therapy according to the inventionmay be administered in the form of the raw chemical compound, it ispreferred to introduce the active ingredient, optionally in the form ofa physiologically acceptable salt, in a pharmaceutical compositiontogether with one or more adjuvants, excipients, carriers, buffers,diluents, and/or other customary pharmaceutical auxiliaries.

In a preferred embodiment, the invention provides pharmaceuticalcompositions comprising the chemical compound for use according to theinvention, or a pharmaceutically acceptable salt or derivative thereof,together with one or more pharmaceutically acceptable carrierstherefore, and, optionally, other therapeutic and/or prophylacticingredients, know and used in the art. The carrier(s) must be“acceptable” in the sense of being compatible with the other ingredientsof the formulation and not harmful to the recipient thereof. In afurther embodiment, the invention provides pharmaceutical compositionscomprising more than one compound or prodrug for use according to theinvention, such as two different compounds or prodrugs for use accordingto the invention.

Pharmaceutical compositions of the invention may be those suitable fororal, rectal, bronchial, nasal, pulmonal, topical (including buccal andsub-lingual), transdermal, vaginal or parenteral (including cutaneous,subcutaneous, intramuscular, intraperitoneal, intravenous,intraarterial, intracerebral, intraocular injection or infusion)administration, or those in a form suitable for administration byinhalation or insufflation, including powders and liquid aerosoladministration, or by sustained release systems. Suitable examples ofsustained release systems include semipermeable matrices of solidhydrophobic polymers containing the compound of the invention, whichmatrices may be in form of shaped articles, e.g. films or microcapsules.

The chemical compound of the invention, together with a conventionaladjuvant, carrier, or diluent, may thus be placed into the form ofpharmaceutical compositions and unit dosages thereof. Such forms includesolids, and in particular tablets, filled capsules, powder and pelletforms, and liquids, in particular aqueous or non-aqueous solutions,suspensions, emulsions, elixirs, and capsules filled with the same, allfor oral use, suppositories for rectal administration, and sterileinjectable solutions for parenteral use. Such pharmaceuticalcompositions and unit dosage forms thereof may comprise conventionalingredients in conventional proportions, with or without additionalactive compounds or principles, and such unit dosage forms may containany suitable effective amount of the active ingredient commensurate withthe intended daily dosage range to be employed.

The chemical compound of the present invention can be administered in awide variety of oral and parenteral dosage forms. It will be obvious tothose skilled in the art that the following dosage forms may comprise,as the active component, either a chemical compound of the invention ora pharmaceutically acceptable salt of a chemical compound of theinvention.

For preparing pharmaceutical compositions from a chemical compound ofthe present invention, pharmaceutically acceptable carriers can beeither solid or liquid. Solid form preparations include powders,tablets, pills, capsules, cachets, suppositories, and dispersiblegranules. A solid carrier can be one or more substances which may alsoact as diluents, flavouring agents, solubilizers, lubricants, suspendingagents, binders, preservatives, tablet disintegrating agents, or anencapsulating material.

In powders, the carrier is a finely divided solid, which is in a mixturewith the finely divided active component.

In tablets, the active component is mixed with the carrier having thenecessary binding capacity in suitable proportions and compacted in theshape and size desired.

The powders and tablets preferably contain from five or ten to aboutseventy percent of the active compound. Suitable carriers are magnesiumcarbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin,starch, gelatin, tragacanth, methylcellulose, sodiumcarboxymethylcellulose, a low melting wax, cocoa butter, and the like.The term “preparation” is intended to include the formulation of theactive compound with encapsulating material as carrier providing acapsule in which the active component, with or without carriers, issurrounded by a carrier, which is thus in association with it.Similarly, cachets and lozenges are included. Tablets, powders,capsules, pills, cachets, and lozenges can be used as solid formssuitable for oral administration.

For preparing suppositories, a low melting wax, such as a mixture offatty acid glyceride or cocoa butter, is first melted and the activecomponent is dispersed homogeneously therein, as by stirring. The moltenhomogenous mixture is then poured into convenient sized moulds, allowedto cool, and thereby to solidify.

Compositions suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or sprays containing inaddition to the active ingredient such carriers as are known in the artto be appropriate.

Liquid preparations include solutions, suspensions, and emulsions, forexample, water or water-propylene glycol solutions. For example,parenteral injection liquid preparations can be formulated as solutionsin aqueous polyethylene glycol solution.

The chemical compound according to the present invention may thus beformulated for parenteral administration (e.g. by injection, for examplebolus injection or continuous infusion) and may be presented in unitdose form in ampoules, pre-filled syringes, small volume infusion or inmulti-dose containers with an added preservative. The compositions maytake such forms as suspensions, solutions, or emulsions in oily oraqueous vehicles, and may contain formulation agents such as suspending,stabilising and/or dispersing agents. Alternatively, the activeingredient may be in powder form, obtained by aseptic isolation ofsterile solid or by lyophilization from solution, for constitution witha suitable vehicle, e.g. sterile, pyrogen-free water, before use.

Aqueous solutions suitable for oral use can be prepared by dissolvingthe active component in water and adding suitable colorants, flavours,stabilising and thickening agents, as desired.

Aqueous suspensions suitable for oral use can be made by dispersing thefinely divided active component in water with viscous material, such asnatural or synthetic gums, resins, methylcellulose, sodiumcarboxymethylcellulose, or other well known suspending agents.

Also included are solid form preparations, intended for conversionshortly before use to liquid form preparations for oral administration.Such liquid forms include solutions, suspensions, and emulsions. Inaddition to the active component such preparations may comprisecolorants, flavours, stabilisers, buffers, artificial and naturalsweeteners, dispersants, thickeners, solubilizing agents, and the like.

For topical administration to the epidermis the chemical compound of theinvention may be formulated as ointments, creams or lotions, or as atransdermal patch. Ointments and creams may, for example, be formulatedwith an aqueous or oily base with the addition of suitable thickeningand/or gelling agents. Lotions may be formulated with an aqueous or oilybase and will in general also contain one or more emulsifying agents,stabilising agents, dispersing agents, suspending agents, thickeningagents, or colouring agents.

Compositions suitable for topical administration in the mouth includelozenges comprising the active agent in a flavoured base, usuallysucrose and acacia or tragacanth; pastilles comprising the activeingredient in an inert base such as gelatin and glycerine or sucrose andacacia; and mouthwashes comprising the active ingredient in a suitableliquid carrier.

Solutions or suspensions are applied directly to the nasal cavity byconventional means, for example with a dropper, pipette or spray. Thecompositions may be provided in single or multi-dose form.

Administration to the respiratory tract may also be achieved by means ofan aerosol formulation in which the active ingredient is provided in apressurised pack with a suitable propellant such as a chlorofluorocarbon(CFC) for example dichlorodifluoromethane, trichlorofluoromethane, ordichlorotetrafluoroethane, carbon dioxide, or other suitable gas. Theaerosol may conveniently also contain a surfactant such as lecithin. Thedose of drug may be controlled by provision of a metered valve.

Alternatively the active ingredients may be provided in the form of adry powder, for example a powder mix of the compound in a suitablepowder base such as lactose, starch, starch derivatives such ashydroxypropylmethyl cellulose and polyvinylpyrrolidone (PVP).Conveniently the powder carrier will form a gel in the nasal cavity. Thepowder composition may be presented in unit dose form for example incapsules or cartridges of, e.g., gelatin, or blister packs from whichthe powder may be administered by means of an inhaler.

In compositions intended for administration to the respiratory tract,including intranasal compositions, the compound will generally have asmall particle size for example of the order of 5 microns or less. Sucha particle size may be obtained by means known in the art, for exampleby micronization.

When desired, compositions adapted to give sustained release of theactive ingredient may be employed.

The pharmaceutical preparations are preferably in unit dosage forms. Insuch form, the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packaged tablets, capsules, and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form.

Tablets or capsules for oral administration and liquids for intravenousadministration and continuous infusion are preferred compositions.

Further details on techniques for formulation and administration may befound in the latest edition of Remington's Pharmaceutical Sciences(Maack Publishing Co, Easton, Pa.).

A therapeutically effective dose refers to that amount of activeingredient, which ameliorates the symptoms or condition. Therapeuticefficacy and toxicity, e.g. ED₅₀ and LD₅₀, may be determined by standardpharmacological procedures in cell cultures or experimental animals. Thedose ratio between therapeutic and toxic effects is the therapeuticindex and may be expressed by the ratio LD₅₀/ED₅₀. Pharmaceuticalcompositions exhibiting large therapeutic indexes are preferred.

The dose administered must of course be carefully adjusted to the age,weight and condition of the individual being treated, as well as theroute of administration, dosage form and regimen, and the resultdesired, and the exact dosage should of course be determined by thepractitioner.

The actual dosage depend on the nature and severity of the disease beingtreated, and is within the discretion of the physician, and may bevaried by titration of the dosage to the particular circumstances ofthis invention to produce the desired therapeutic effect. However, it ispresently contemplated that pharmaceutical compositions containing offrom about 0.1 to about 1000 mg of active ingredient per individualdose, preferably of from about 1 to about 100 mg, are suitable fortherapeutic treatments.

The active ingredient may be administered in one or several doses perday. Preferred ranges are from 10-200 mg/day p.o. administered in one ortwo doses, such as from 25-50 mg p.o. twice a day.

Ophthalmic Formulations

The pharmaceutical composition may be prepared in unit dosage formssuitable for topical ocular use. The therapeutically efficient amounttypically is between 0.0001 and 5% (w/v), preferably between 0.001 and1.0% (w/v) in liquid formulations.

For ophthalmic application, preferably solutions are prepared using aphysiological saline solution as a major vehicle. The pH of suchophthalmic solutions should preferably be maintained between 4.5 and8.0, more preferably between 6.5 and 7.2, with an appropriate buffersystem. The formulations may also contain conventional, pharmaceuticallyacceptable preservatives, stabilizers and surfactants.

The preservative may be selected from hydrophobic or non-ionicpreservatives, anionic preservatives, and cationic preservatives.Preferred preservatives that may be used in the pharmaceuticalcompositions of the present invention include, but are not limited to,benzalkonium chloride, chlorobutanol, thimerosal, phenylmercuric acetateand phenylmercuric nitrate.

A preferred surfactant is, for example, Polysorbate 80. Likewise,various preferred vehicles may be used in the ophthalmic preparations ofthe present invention. These vehicles include, but are not limited to,polyvinyl alcohol, povidone, hydroxypropyl methyl cellulose, poloxamers,carboxymethyl cellulose, hydroxyethyl cellulose and purified water.

Tonicity adjustors, such as non-ionic tonicity adjustors, may be addedas needed or convenient. They include, but are not limited to, salts,particularly sodium chloride, potassium chloride, mannitol and glycerol,polyethylene glycols (PEG), polypropylene glucols (PPG) or any othersuitable ophthalmically acceptable tonicity adjustor.

Various buffers and means for adjusting pH may be used so long as theresulting preparation is ophthalmically acceptable. Accordingly, buffersinclude acetate buffers, citrate buffers, phosphate buffers and boratebuffers. Acids or bases may be used to adjust the pH of theseformulations as needed.

An ophthalmically acceptable antioxidant for use in the presentinvention includes, but is not limited to, sodium metabisulfite, sodiumthiosulfate, acetylcysteine, butylated hydroxyanisole and butylatedhydroxytoluene.

Other excipient components which may be included in the ophthalmicpreparations are chelating agents. The preferred chelating agent isedentate disodium, although other chelating agents may also be used inplace or in conjunction with it.

The ingredients are usually used in the following amounts: IngredientAmount (% w/v) Active compound 0.001-5 Preservative    0-0.10 Vehicle   0-40 Tonicity adjustor    1-10 Buffer  0.01-10 pH adjustor q.s. pH4.5-8.0 Antioxidant as needed Surfactant as needed Purified water asneeded to make 100%

The actual dose of the active compounds of the present invention dependson the specific compound, and on the condition to be treated; theselection of the appropriate dose is well within the knowledge of theskilled artisan.

The ophthalmic formulations of the present invention are convenientlypackaged in forms suitable for metered application, such as incontainers equipped with a dropper, to facilitate the application to theeye. Containers suitable for dropwise application are usually made ofsuitable inert, non-toxic plastic material, and generally containbetween about 0.5 and about 15 ml solution.

In the case of treating ophthalmic angiogenesis related diseases,disorders or conditions—such as AMD, the pharmaceutical composition ofthe invention may also be administered in the form of systemicadministration (such as orally), as an eye ointment, or as an injectionin the eye (periocular or intraocular injection).

Methods of Therapy

The invention also provides a method for the treatment, prevention oralleviation of a disease or a disorder or a condition of a living animalbody, including a human, which disease, disorder or condition isresponsive to inhibition of angiogenesis, and which method comprisesadministering to such a living animal body, including a human, in needthereof an effective amount of a compound of general formula I asdefined above.

The diseases, disorders or conditions that are responsive to inhibitionof angiogenesis include but are not limited to:

-   -   diseases, disorders or conditions that involve the proliferation        of tumor cells, such as cancer, prostate cancer, lung cancer,        breast cancer, bladder cancer, renal cancer, colon cancer,        gastric cancer, pancreatic cancer, ovarian cancer, melanoma,        hepatoma, sarcoma and lymphoma;    -   ophthalmic angiogenesis related diseases, disorders or        conditions, such as exudative macular degeneration, age-related        macular degeneration (AMD), retinopathy, diabetic retinopathy,        proliferative diabetic retinopathy, diabetic macular edema        (DME), ischemic retinopathy (e.g. retinal vain or artery        occlusion), retinopathy of prematurity, neovascular glaucoma,        and corneal neovascularization; and    -   rheumatoid arthritis, and psoriasis.

In a special embodiment, the disease, disorder or condition to betreated is a preneoplastic disease state. In a further embodiment, thetreatment is an anti-metastatic treatment. In a still furtherembodiment, the disease, disorder or condition to be prevented ismetastatic cancer. In a further embodiment, the disease, disorder orcondition to be prevented or alleviated is DME.

Also the invention provides a method of treatment, prevention oralleviation of age-related macular degeneration of a living animal body,including a human, which method comprises administering to such a livinganimal body, including a human, in need thereof a therapeuticallyeffective amount of a VRAC blocker or a pharmaceutically acceptable saltthereof.

In the context of this invention, “age-related macular degeneration”(AMD) includes dry AMD (non-exudative AMD) and wet AMD (exudative AMD).

In a special embodiment, the invention relates to treatment, preventionor alleviation of wet AMD.

Combined Therapy

The pharmaceutical composition for use according to the invention mayinclude or may be used or administered in combination with one or moreadditional drugs useful for the treatment, prevention or alleviation ofa disease responsive to inhibition of angiogenesis, such as compoundsuseful for anti-metastatic treatment. Such additional drugs includecytotoxic compounds, antimitotic compounds, and antimetabolites.

Examples of cytotoxic compounds (including cytotoxic alkylating agents)include carmustine (BCNU), fotemustin, temozolomide (temodal),ifosfamide, and cyclofosfamide.

Examples of antimitotic compounds include paclitaxel (taxol) anddocetaxel.

An example of antimetabolites includes methotrexat.

Furthermore, the pharmaceutical composition for use according to theinvention may be used or administered in combination with othertreatments or therapies. Examples of other treatments or therapiesinclude radiotherapy and surgery.

Test Methods

The efficacy of use of the compound according to the invention may beevaluated by standard in vitro and in vivo studies as e.g. thosedescribed below.

In vitro Methods

Cell-Specificity Assay: Incorporation of [³H]Thymidine

Confluent cultures of HUVEC, fibroblasts, Mel 57 and T47D cells weredetached by trypsin/EDTA solution, and allowed to adhere and spread atan appropriate cell density on gelatine-coated dishes in M199-HEPESmedium or DMEM-HEPES medium both supplemented with 10% heat-inactivatednew born calve serum (NBCS) and penidillin/streptomycin. After 18 h theHUVEC and fibroblasts were stimulated with 2.5 ng/ml FGF-2 inM199-HEPES, penicillin/streptomycin, 10% NBCS and 0.1% DMSO in duplicatewells, with or without the test compounds. The tumor cells were culturedin DMEM-HEPES supplemented with 10% NBCS, penicillin/streptomycin and0.1% DMSO in duplicate wells, with or without the test compounds. Afteran incubation period of 48 h, a tracer amount (0.5 μCi/well) of[³H]thymidine was added and the cells were incubated for another 6 hperiod. Subsequently, the cells were washed with PBS, [3H]-labelled DNAwere fixed with methanol, and precipitated in 5% trichloroacetic-acid,and finally dissolved in 0.5 ml 0.3 M NaOH and counted in a liquidscintillation counter.

Cell Morphology and Proliferation Assay

One week before the assay, a vial with HUVEC (passage 1) was thawed andcultured (after a split ratio of 1:3) to confluence (passage 2). Theconfluent culture of HUVEC was detached by trypsin/EDTA solution, andallowed to adhere and spread at cell density of 10, 50 and 100%confluency on gelatine-coated dishes in M199-HEPES medium supplementedwith 10% heat-inactivated NBCS, 10% human serum andpenicillin/streptomycin. After 18 h the HUVEC were preincubated with thetest compounds for 4 hours. Then the HUVEC were washed and restimulatedin the absence or presence of the test compounds and the referencecompounds with 2.5 ng/ml FGF-2 in M199-HEPES, penicillin/streptomycin,10% NBCS, 10% human serum and 0.1% DMSO in triplicate wells for 5 (10%confluency) or 3 days (50 and 100% confluency), with or without the testcompounds. The cell number was determined by image analysis (P.Koolwijk, 2001)

Observations, Analyses and Measurements

All outcome measures were measured in singular, i.e. one measurement perculture well. The proliferation of HUVEC, fibroblast, Mel 57, and T47Dtumor cells was expressed as mean±range [3H]-thymidine incorporation(dpm) of duplicate wells.

The percentage of inhibition of FGF-2-induced HUVEC and fibroblastproliferation by the compounds was calculated as follows:${\%\quad{inhibition}} = {\left( {1 - \frac{\left( {\left( {{dpm},{HUVEC}_{{FGF} - 2}} \right) - \left( {{dpm},{HUVEC}_{{FGF} - 2 + {compound}}} \right)} \right)}{\left( {\left( {{dppm},{HUVEC}_{{FGF} - 2}} \right) - \left( {{dpm},{HUVEC}_{control}} \right)} \right)}} \right) \times 100\%}$HUVEC_(control) = non  stimulated    HUVECHUVEC_(FGF − 2) = FGF − 2  stimulated    HUVECHUVEC_(FGF − 2 + compound) = FGF − 2  stimulated    HUVEC + test  compundThe percentage of inhibition of the Mel 57 and T47D tumor cellproliferation by the compounds will be calculated as follows:${\%\quad{inhibition}} = {\left( {1 - \frac{\left( {\left( {{dpm},{{tumor}\quad{cell}}} \right) - \left( {{dpm},{{tumor}{\quad\quad}{cell}_{+ {compund}}}} \right)} \right)}{\left( {{dpm},{{tumor}\quad{cell}}} \right)}} \right) \times 100\%}$tumor  cell = NBCS  stimulated  tumor  cell  tumor  cell_(+compound) = NBCS  stimulated  tumor    cell + test  compoundIn vivo MethodsMouse Anti-Angiogenesis Assay

NMRI female mice (SPF Bom:NMRI) weighing 25-27 g were obtained from M&B,Ejby, Lille Skensved, Denmark. They were housed in a facility wherelight was controlled on a 12 hour light-dark cycle. The room temperatureand relative humidity recorded by a thermo-hygrograph showed valuesbetween 20.5-24.1° C. and 40-67%, respectively. The animals were fed apelleted rodent diet (Altromin 1324, Brogarden, Denmark) ad libitum andhad free access to tap water. All animals were observed daily forclinical signs.

Slow-release pellets containing 400 ng of human basic fibroblast growthfactor (Innovative Research of America, Florida, USA) were circular andwith a diameter of 1.5 mm. The angiogenic peptide was guarantied by thesupplier to be released over a period of 10 days.

The mice were anaesthetized using inhalation anaesthesia (halothane/N₂Oand oxygen). The skin of the back was shaved using an electric shaverand the skin was disinfected using 70% ethanol. A 5 mm incision was madetransversely in close proximity to the shoulder blades and a 2 cm pocketreaching caudally to the pelvic region was created by blunt dissection,carefully separating the skin from the fascie. A polyurethane spongewith the dimension of 8×5×3 mm containing a slow-release pellet of 400ng bFGF was placed at the caudal end of the pocket and the incision wasclosed by a single or double invert suture using Perma-Hand Seide 4/0(Johnson & Johnson, Brussels, Belgium). The animals were treated by ananalgesic subcutaneous injection of carprofen 2 mg/kg.

The angiogenic response was quantitated as previously described(Lichtenberg et al., 1997, 1999 & 2002). Briefly, twenty minutes beforeeuthanasia 1 μCi of ¹²⁵I-labelled immuno-globulin (Amersham, UK) in 50μl of 0.9% NaCl was injected intravenously into a tail vein. The animalswere euthanised by O₂/CO₂ asphyxiation and the skin overlying the spongeimplant was removed. The sponge implant with the pellet was placed in aplastic vial containing 4% formalin and the ¹²⁵I-activity was measuredin a γ-counter. Differences in angiogenic response measured as¹²⁵I-activity in cpm were assessed by Student's t-test, grouped data,with P<0.05 regarded as statistically significant. Data was expressed asmeans±SEM.

Mouse Metastasis Assay

Female C57BL/6 mice were supplied and delivered by Charles River UK Ltd.The animals were approximately 6 weeks of age at the start of the study.The body weights at the start of dosing were in the range 10-21 g. Themice were housed in solid-bottomed plastic cages, containing woodshavings, in groups of up to 10. During acclimatization, the rooms andcages were cleaned at regular intervals to maintain hygiene. The micewere fed an expanded rodent diet ad libitum and allowed free access tomains tap water. The holding rooms had a 12 h light-dark cycle, and wereair-conditioned by a system designed to maintain air temperature withinthe range 20±3° C. (McKay, 2002).

Data was expressed as means±SEM and analysed using appropriatestatistical methods. Statistical significance was assumed when P<0.05.

BRIEF DESCRIPTION OF THE DRAWINGS

The present invention is further illustrated by reference to theaccompanying drawing, in which:

FIG. 1 shows the effects of Compound a on HUVEC (◯), fibroblast (▴) andtumor cell (Mel 57(●) and T47D(∇)) proliferation;

FIG. 2 shows the effects of Compound aon 10%, 50% and 100% confluentmonolayers of HUVEC. Solid symbols and bars: continuous conditions; opensymbols and bars: preincubated conditions.

EXAMPLES

The invention is further illustrated with reference to the followingexamples, which are not intended to be in any way limiting to the scopeof the invention as claimed. The examples describe test results for thecompounds N-4-Nitrophenyl-N′-[4-bromo-2-(1-H-tetrazol-5-yl)phenyl]urea(compound a) andN-3,5-Di(trifluoromethyl)phenyl-N′-[4-bromo-2-(1-H-tetrazol-5-yl)phenyl]urea(compound b).

Example 1

Compound a Tested in the in vitro Cell Specificity Assay

A difference in cell-specificity was observed of Compound a (see FIG.1). The compound was a more potent inhibitor of the bFGF-induced HUVECand fibroblast proliferation when compared to the NBCS-induced Mel 57and T47D proliferation.

Example 2

Compound a Tested in the in vitro Cell Morphology and ProliferationAssay

To investigate the mode of action of the compound on proliferating andnon-proliferating HUVEC, experiments were performed on 10%, 50% and 100%confluent monolayers of HUVEC. The 10% and 50% confluent HUVECmonolayers represent the status of angiogenic endothelial cells able toproliferate and migrate when stimulated. The 100% HUVEC monolayersrepresent the quiescent character of the endothelial cells in theexisting blood vessel. The 4-hour preincubation experiments wereperformed to be able to discriminate between general toxicity and theinduction of apoptosis.

Compound a was not able to inhibit HUVEC proliferation significantlywhen added for the 4-hour preincubation period and then removed for therest of the stimulation period (see FIG. 2). No signs of anycytotoxicity (cell death indicated by the observation of floating cellsin the media) was observed during this period. In addition, there was nodifference in cell death or any delay of cell growth by thepreincubation period during the rest of the 3-day or 5day incubationperiod.

However, when the incubation was performed in the continuous presence ofthe compound, there was a clear inhibitory effect on the proliferationof the cells at the 10% confluence HUVEC monolayers. This effect wasobserved at the two highest concentrations of the compound, but not atlower concentrations. The compound induced inhibition of HUVECproliferation was also slightly observed at the 50% confluent monolayers(see FIG. 2). There was no effect of the compound on the 100% HUVECmonolayers.

This non-cytotoxicity of the compound on the 100% confluent monolayerswas confirmed by the fact that there was no change of HUVEC morphologyand the amount of floating dead cells in the culture medium observedduring the culture periods with the compound.

Example 3

Compounds a and b Tested in the in vivo Mouse Anti-Angiogenesis Assay

Three separate experiments were conducted: Two experiments wereperformed with Compound a and one experiment was performed with Compoundb. In the first two experiments with Compound a, 3 groups eachcomprising 5 animals was treated orally with Compound a at the doselevels of 0 (saline), 5 and 10 mg/kg/day (experiment 1) and with 20 and40 mg/kg/day (experiment 2). In the third experiment, 3 groups eachcomprising 6 animals was treated orally with the vehicle (saline) and 80mg/kg/day of Compound a. In all experiments the animals were treatedfrom Days 3-9 and killed at day 10.

Compound a elicited a dose-response relationship at all doses. At 10 and40 mg/kg/day a significant inhibition of the neovacularisation of 37%and 48% was obtained (see Table 1). A dose of 5 mglkg/day appeared to bethe No Effect Level (NOEL).

Compound b at a dose of 80 mg/kg/day inhibited the angiogenesis responseof approximately 60% compared to vehicle treated animals. The selecteddose levels were well tolerated by the mice; no signs of toxicity orchanges of the body weight gain were observed (data not shown). TABLE 1The effect of Compounds a and b on the neovascularisation in mice(expressed as percent inhibition) Dose mg/kg Treatment 5 10 20 40 80Compound a 15% 37%* 28% 48%* Compound b 61%**P < 0.05, compared to vehicle (t-test)

Example 4

Compounds a and b Tested in the in vivo Mouse Metastasis Assay

Two separate studies were conducted: One was performed with Compound a(experiment 1) and one with Compound b (experiment 2). In each studythere were 4 treatment groups. The treatment groups were as follows:Group Treatment Dose 1 Untreated Control — 2 Vehicle Control 20 ml/kgp.o. 3 Compounds a or b 60 mg/kg p.o. 4 Compounds a or b 80 mg/kg p.o.

Treatments for groups 24 were administered orally, by gavage. The dosevolume used was 20 ml/kg for groups 2 and 4 and 15 ml/kg for group 3.

C57BU6 mice were injected intravenously, via a tail vein, with 0.1 ml ofa suspension of B16/F10 melanoma cells (approximately 3×10⁵ cells/mouse)on day 0. With the exception of the untreated group, animals were dosedorally, by gavage, or intravenously, according to their allocatedtreatment group once daily, from day −2 to day 10 (13 administrations).The animals were killed on day 14 (14 days after injection of the tumourcells). The lungs from each animal were removed and weighed prior tofixing in Bouin's solution. After fixation, the number of colonies onthe surface of each set of lungs was counted by eye and these data wereused for statistical analysis.

Oral administration of Compound a, at doses of 60 and 80 mg/kg, resultedin a significant reduction of 17% and 21%, respectively in the number ofmelanoma lung colonies when compared to vehicle treated mice (see Table2).

Oral administration of Compound b, at similar doses (60 and 80 mg/kg),produced a significant reduction of 36% and 44%, respectively in thenumber of melanoma lung colonies compared to vehicle treated mice (seeTable 3). TABLE 2 The effect of Compound a on the development of B16melanoma lung colonies in C57BL/6 mice Group Treatment Colony Count %Reduction 1 Untreated Control — — 2 Vehicle Control (20 ml/kg) 78.29 ±3.56 — 3 Compound a (60 mg/kg) 64.68 ± 2.65* 17.38 4 Compound a (80mg/kg) 61.85 ± 2.89* 21.00*P < 0.01, compared to vehicle (Kruskal-Wallis and Dunnett's test)

TABLE 3 The effect of Compound b on the development of B16 melanoma lungcolonies in C57BL/6 mice Group Treatment Colony Count % Reduction 1Untreated Control — — 2 Vehicle Control (20 ml/kg) 65.40 ± 7.90 — 3Compound b (60 mg/kg) 41.55 ± 9.55* 36.45 4 Compound b (80 mg/kg) 36.65± 5.82* 43.97*P < 0.01, compared to vehicle (Kruskal-Wallis and Dunnett's test)

1-11. (canceled)
 12. A method of treatment, prevention or alleviation ofa disease or a disorder or a condition of a living animal body,including a human, which disorder, disease or condition is responsive toinhibition of angiogenesis, comprising the step of administering to sucha living animal body, including a human, in need thereof atherapeutically effective amount of a compound of general formula I

or a pharmaceutically acceptable salt thereof wherein R² representstetrazolyl; R¹, R⁴, R⁵, R⁶, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ independently ofeach other represent hydrogen, halo, trifluoromethyl, nitro, alkyl,alkylcarbonyl, —NR^(a)R^(b), —NR^(a)—CO—R^(b), phenyl or heteroaryl;which phenyl is optionally substituted with halo, trifluoromethyl,nitro, —CO—NHR^(c), —CO—O—R^(c) or —CO—N′R″; wherein R^(c) is hydrogen,alkyl, or phenyl; R′ and R″ independently of each other are hydrogen oralkyl; or R′ and R″ together with the nitrogen to which they areattached form a 5- to 7-membered heterocyclic ring, which ring mayoptionally comprise as a ring member, one oxygen atom, and/or oneadditional nitrogen atom, and/or one carbon-carbon double bond, and/orone carbon-nitrogen double bond; and which heterocyclic ring mayoptionally be substituted with alkyl; R^(a) and R^(b) independently ofeach other are hydrogen or alkyl; or R¹⁵ and R¹⁶, or R¹⁴ and R¹⁵together with the phenyl ring to which they are attached form a naphthylring or an indanyl ring; and R³, R⁴, R⁵, R⁶, R¹² and R¹³ and theremaining one of R¹⁴, R¹⁵ and R¹⁶ are as defined above.
 13. The methodaccording to claim 12, wherein R³, R⁵, and R⁶ represent hydrogen; and R⁴represents halo.
 14. The method according to claim 12, wherein R³, R⁵,and R⁶ represent hydrogen; and R⁴ represents phenyl substituted withtrifluoromethyl, nitro or —CO—NHR^(c); wherein R^(c) is phenyl.
 15. Themethod according to claim 12, wherein the compound isN-4-Nitrophenyl-N′-[4-bromo-2-(1-H-tetrazol-5-yl)phenyl]urea;N-3,5-Di(trifluoromethyl)phenyl-N′-[4-bromo-2-(1-H-tetrazol-5-yl)phenyl]urea;N-3-Trifluoromethylphenyl-N′-[4-(3-nitrophenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;N-3-Trifluoromethylphenyl-N′-[4-(4-anilinocarbonylphenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;N-3-Trifluoromethylphenyl-N′-[4-(4-trifluoromethylphenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;N-(3-Trifluoromethyl-phenyl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Trifluoromethyl-phenyl)-N′-[4-bromo-2-(1-H-terazol-5-yl)-phenyl]urea;N-(3-Trifluoromethyl-phenyl)-N′-[4-phenyl-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Chloro-phenyl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Trifluoromethyl-phenyl)-N′-[4-amino-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Trifluoromethyl-phenyl)-N′-[4-acetylamino-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Trifluoromethyl-phenyl)-N′-[4-carbamoyl-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Trifluoromethyl-phenyl)-N′-[4-(N″,N″-dimethylcarbamoyl)-2-(1-H-tetrazol-5-yl)-phenyl]urea;3′-(1-H-tetrazol-5-yl)-4′-[3-(3-trifluoromethyl-phenyl)-ureido]-biphenyl-4-carboxylicacid; -p0 N-(Indan-5-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(Biphenyl-4-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(Biphenyl-3-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Acetyl-phenyl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(Biphenyl-3-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-[3-(Pyridin-3-yl)-phenyl]-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Bromo-phenyl)-N′-[4′-(4-methyl-piperazine-1-carbonyl)-3-(1-H-tetrazol-5-yl)-biphenyl-4-yl]urea;N-(3,5-Dichloro-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3,4-Dichloro-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(Naphthalen-1-yl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(2-Trifluoromethyl-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(2-Fluoro-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(2-Ethyl-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea; or apharmaceutically acceptable salt thereof.
 16. The method according toclaim 12, wherein the disease, disorder or condition that is responsiveto inhibition of angiogenesis is selected from the group consisting ofcancer, prostate cancer, lung cancer, breast cancer, bladder cancer,renal cancer, colon cancer, gastric cancer, pancreatic cancer, ovariancancer, melanoma, hepatoma, sarcoma, lymphoma, exudative maculardegeneration, age-related macular degeneration, retinopathy, diabeticretinopathy, proliferative diabetic retinopathy, diabetic macular edema(DME), ischemic retinopathy, retinopathy of prematurity, neovascularglaucoma, corneal neovascularization, rheumatoid arthritis, andpsoriasis.
 17. The method according to claim 12, wherein the compound isN-4-Nitrophenyl-N′-[4-bromo-2-(1-H-tetrazol-5-yl)phenyl]urea;N-3,5-Di(trifluoromethyl)phenyl-N′-[4-bromo-2-(1-H-tetrazol-5-yl)phenyl]urea;N-3-Trifluoromethylphenyl-N′-[4-(3-nitrophenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;N-3-Trifluoromethylphenyl-N′-[4-(4-anilinocarbonylphenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;N-3-Trifluoromethylphenyl-N′-[4-(4-trifluoromethylphenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;or a pharmaceutically acceptable salt thereof, and the treatment is ananti-metastatic treatment.
 18. A method of treatment, prevention oralleviation of age-related macular degeneration of a living animal body,including a human comprising the step of administering to such a livinganimal body, including a human, in need thereof a therapeuticallyeffective amount of a VRAC blocker or a pharmaceutically acceptable saltthereof.
 19. The method according to 18, wherein the VRAC blocker is acompound of general formula I

or a pharmaceutically acceptable salt thereof wherein R² representstetrazolyl; and R³, R⁴, R⁵, R⁶, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶independently of each other represent hydrogen, halo, trifluoromethyl,nitro, alkyl, alkylcarbonyl, —NR^(a-l R) ^(b), NR^(a)—CO—R^(b), phenylor heteroaryl; which phenyl is optionally substituted with halo,trifluoromethyl, nitro, —CO—NHR^(c), —CO—O—R^(c) or —CO—NR′R″; whereinR^(c) is hydrogen, alkyl, or phenyl; R′ and R″ independently of eachother are hydrogen or alkyl; or R′ and R″ together with the nitrogen towhich they are attached form a 5- to 7-membered heterocyclic ring, whichring may optionally comprise as a ring member, one oxygen atom, and/orone additional nitrogen atom, and/or one carbon-carbon double bond,and/or one carbon-nitrogen double bond; and which heterocyclic ring mayoptionally be substituted with alkyl; R^(a) and R^(b) independently ofeach other are hydrogen or alkyl; or R¹⁵ and R¹⁶, or R¹⁴ and R¹⁵together with the phenyl ring to which they are attached form a naphthylring or an indanyl ring; and R³, R⁴, R⁵, R⁶, R¹² and R¹³ and theremaining one of R¹⁴, R¹⁵ and R¹⁶ are as defined above.
 20. The methodaccording to claim 18, wherein the compound isN-4-Nitrophenyl-N′-[4-bromo-2-(1-H-tetrazol-5-yl)phenyl]urea;N-3,5-Di(trifluoromethyl)phenyl-N′-[4-bromo-2-(1-H-tetrazol-5-yl)phenyl]urea;N-3-Trifluoromethylphenyl-N′-[4-(3-nitrophenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;N-3-Trifluoromethylphenyl-N′-[4-(4-anilinocarbonylphenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;N-3-Trifluoromethylphenyl-N′-[4-(4-trifluoromethylphenyl)-2-(1-H-tetrazol-5-yl)phenyl]urea;N-(3-Trifluoromethyl-phenyl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Trifluoromethyl-phenyl)-N′-[4-bromo-2-(1-H-terazol-5-yl)-phenyl]urea;N-(3-Trifluoromethyl-phenyl)-N′-[4-phenyl-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Chloro-phenyl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Trifluoromethyl-phenyl)-N′-[4-amino-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Trifluoromethyl-phenyl)-N′-[4-acetylamino-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Trifluoromethyl-phenyl)-N′-[4-carbamoyl-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Trifluoromethyl-phenyl)-N′-[4-(N″,N″-dimethylcarbamoyl)-2-(1-H-tetrazol-5-yl)-phenyl]urea;3′-(1-H-tetrazol-5-yl)-4′-[3-(3-trifluoromethyl-phenyl)-ureido]-biphenyl-4-carboxylicacid; N-(Indan-5-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(Biphenyl-4-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(Biphenyl-3-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Acetyl-phenyl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(Biphenyl-3-yl)-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-[3-(Pyridin-3-yl)-phenyl]-N′-[2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3-Bromo-phenyl)-N′-[4′-(4-methyl-piperazine-1-carbonyl)-3-(1-H-tetrazol-5-yl)-biphenyl-4-yl]urea;N-(3,5-Dichloro-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(3,4-Dichloro-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(Naphthalen-1-yl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(2-Trifluoromethyl-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(2-Fluoro-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea;N-(2-Ethyl-phenyl)-N′-[4-bromo-2-(1-H-tetrazol-5-yl)-phenyl]urea; or apharmaceutically acceptable salt thereof.